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Simultaneous measurement of NH and CαHα coupling constants in proteins

✍ Scribed by David Pantoja-Uceda; Jorge Santoro

Publisher
John Wiley and Sons
Year
2009
Tongue
English
Weight
241 KB
Volume
48
Category
Article
ISSN
0749-1581

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✦ Synopsis

Abstract

We present a pulse sequence for the simultaneous measurement of NH and CαHα couplings in double‐labeled proteins from 2D spectra. The proposed sequence, a modification of the HN(CO)CA experiment, combines the J‐modulation method and the IPAP scheme. The couplings can be readily retrieved from a series of 2D ^15^N^1^H correlation spectra, differing in the time point at which a ^1^H 180° pulse is applied. This induces an intensity modulation of the ^15^N^1^H correlation peaks with the CαHα coupling. The CαHα coupling is then obtained by fitting the observed intensities to the modulation equation. The NH coupling is measured in each member of the set from peak‐to‐peak separations in the IPAP subspectra. The pulse sequence is experimentally verified with a sample of ^15^N/^13^C‐enriched ubiquitin. Copyright © 2009 John Wiley & Sons, Ltd.

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